Dataset Information

Exploration of CRISPR/Cas9-based gene editing as therapy for osteoarthritis.

ABSTRACT: OBJECTIVES:Osteoarthritis (OA) is a painful and debilitating disease and it is associated with aberrant upregulation of multiple factors, including matrix metalloproteinase 13 (MMP13), interleukin-1? (IL-1?) and nerve growth factor (NGF). In this study, we aimed to use the CRISPR/Cas9 technology, a highly efficient gene-editing tool, to study whether the ablation of OA-associated genes has OA-modifying effects. METHODS:We performed intra-articular injection of adeno-associated virus, which expressed CRISPR/Cas9 components to target each of the genes encoding MMP13, IL-1? and NGF, in a surgically induced OA mouse model. We also tested triple ablations of NGF, MMP13 and IL-1?. RESULTS:Loss-of-function of NGF palliates pain but worsens joint damage in the surgically induced OA model. Ablation of MMP13 or IL-1? reduces the expression of cartilage-degrading enzymes and attenuates structural deterioration. Targeting both MMP13 and IL-1? significantly mitigates the adverse effects of NGF blockade on the joints. CONCLUSIONS:CRISPR-mediated ablation of NGF alleviates OA pain, and deletion of MMP13-1? or IL-1? attenuates structural damage in a post-traumatic OA model. Multiplex ablations of NGF, MMP13 and IL-1? provide benefits on both pain management and joint structure maintenance. Our results suggest that CRISPR-based gene editing is useful for the identification of promising drug targets and the development of feasible therapeutic strategies for OA treatment.

SUBMITTER: Zhao L

PROVIDER: S-EPMC6621547 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Exploration of CRISPR/Cas9-based gene editing as therapy for osteoarthritis.

Zhao Lan L Huang Jian J Fan Yunshan Y Li Jun J You Tianming T He Shisheng S Xiao Guozhi G Chen Di D

Annals of the rheumatic diseases 20190306 5

<h4>Objectives</h4>Osteoarthritis (OA) is a painful and debilitating disease and it is associated with aberrant upregulation of multiple factors, including matrix metalloproteinase 13 (MMP13), interleukin-1β (IL-1β) and nerve growth factor (NGF). In this study, we aimed to use the CRISPR/Cas9 technology, a highly efficient gene-editing tool, to study whether the ablation of OA-associated genes has OA-modifying effects.<h4>Methods</h4>We performed intra-articular injection of adeno-associated vir ...[more]

PMID: 30842121

Similar Datasets

Project description:Many Candida species that cause infection have diploid genomes and do not undergo classical meiosis. The application of clustered regularly interspaced short palindromic repeat-Cas9 (CRISPR-Cas9) gene editing systems has therefore greatly facilitated the generation of gene disruptions and the introduction of specific polymorphisms. However, CRISPR methods are not yet available for all Candida species. We describe here an adaption of a previously developed CRISPR system in Candida parapsilosis that uses an autonomously replicating plasmid. Guide RNAs can be introduced in a single cloning step and are released by cleavage between a tRNA and a ribozyme. The plasmid also contains CAS9 and a selectable nourseothricin SAT1 marker. It can be used for markerless editing in C. parapsilosis, C. orthopsilosis, and C. metapsilosis We also show that CRISPR can easily be used to introduce molecular barcodes and to reintroduce wild-type sequences into edited strains. Heterozygous mutations can be generated, either by careful selection of the distance between the polymorphism and the Cas9 cut site or by providing two different repair templates at the same time. In addition, we have constructed a different autonomously replicating plasmid for CRISPR-Cas9 editing in Candida tropicalis We show that editing can easily be carried out in multiple C. tropicalis isolates. Nonhomologous end joining (NHEJ) repair occurs at a high level in C. metapsilosis and C. tropicalis IMPORTANCE Candida species are a major cause of infection worldwide. The species associated with infection vary with geographical location and with patient population. Infection with Candida tropicalis is particularly common in South America and Asia, and Candida parapsilosis infections are more common in the very young. Molecular methods for manipulating the genomes of these species are still lacking. We describe a simple and efficient CRISPR-based gene editing system that can be applied in the C. parapsilosis species group, including the sister species Candida orthopsilosis and Candida metapsilosis We have also constructed a separate system for gene editing in C. tropicalis.

Dataset Information

Exploration of CRISPR/Cas9-based gene editing as therapy for osteoarthritis.

Publications

Exploration of CRISPR/Cas9-based gene editing as therapy for osteoarthritis.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets